RAF265-CHIR-265-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemERAF265Cat. No.: HY-10248CAS No.: 927880-90-8Synonyms: CHIR-265分式: CHFNO分量: 518.41作靶点: Raf; VEGFR; Autophagy作通路: MAPK/ERK Pathway; Protein Tyrosine Kinase/RTK;Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20

2、C 1 month溶解性数据体外实验 DMSO : 26 mg/mL (50.15 mM)Ethanol : 10 mg/mL (19.29 mM; Need ultrasonic)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 1.9290 mL 9.6449 mL 19.2898 mL5 mM 0.3858 mL 1.9290 mL 3.8580 mL10 mM 0.1929 mL 0.9645 mL 1.9290 mL请根据产品在不同溶剂中的溶解度，选择合

3、适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% EtOH 40% PEG300 5% Tween-80 45% salineSolubility: 1 mg/mL (1.93 mM); Clear solution1/3 Master of Small Molecules 您边的抑制剂师www.MedChe

4、mE2. 请依序添加每种溶剂： 10% EtOH 90% (20% SBE-CD in saline)Solubility: 1 mg/mL (1.93 mM); Suspended solution; Need ultrasonic3. 请依序添加每种溶剂： 10% EtOH 90% corn oilSolubility: 1 mg/mL (1.93 mM); Clear solution; Need warmingBIOLOGICAL ACTIVITY物活性 RAF265种有效的 RAF/VEGFR2 抑制剂。IC50 & Target VEGFR2 RAF体外研究 The MTT ass

5、ay reveals that in HT29 and MDAMB231 cells, RAF265 alone shows significant activity withIC20 values of 1 to 3 M and IC50 values of 5 to 10 M. In A549 and HCT116 cells, IC20 values are 1 M forboth, but RAF265 concentrations up to 10 M do not reach IC50 values. However, in the presence of 1 nMRAD001,

6、the IC50 for RAF265 is 5 M in A549 cells and 10 M in HCT116 cells 1.体内研究 In single-compound efficacy studies, optimal dosing of RAD001 and RAF265 is 5 to 12 mg/kg daily and 30mg/kg every two days, respectively. However, combination tolerability studies in nontumor-bearing mice defindose-limiting tox

7、icity as a 10% weight loss with the combination of RAD001 at a dose of 12 mg/kg daily andRAF265 at a dose of 20 mg/kg every two days. Therefore, the combination of RAF265 at a dose of 12 mg/kgqd and RAD001 at a dose of 12 mg/kg qd seems to be the maximal tolerated dose. RAD001 and RAF265are both giv

8、en at a dose of 12 mg/kg qd, alone or concurrently, over 6 days. After a 2-day stop, thecompounds are given for another 6 days, and the treatment is then stopped. To confirm the potential of thecombination of RAF265 and RAD001, the antitumor effect of the combination is tested in HCT116 xenografts(K

9、RAS mut, PIK3CA mut). In HCT116 xenografts, RAD001 or RAF265 given alone shows 60% to 65% and71% to 72% TVI%, respectively 1.PROTOCOLCell Assay 1 The MTT assay and Bliss additivism model are used to assess the effect of the combination on cell viability.Human A549 and H460 lung, HT29 and HCT 116 col

10、on, and MDAMB231 breast cancer cell lines are used.In each well of a 96-well plate, 1104 cells are grown in 200 L of medium. After 24 h, RAD001, RAF265, orthe combination is added to achieve a final concentration of 0.1 to 10 nM and 0.1 to 10 M, respectively.After 48 h of treatment, 20 L of 5 mg/mL

11、MTT solution in PBS is added to each well. After 4 h, supernatantis removed and formazan crystals are discarded in 200 L of DMSO. Absorbance is then measured at 595nm using an absorbance plate reader. Data are expressed as the percentage of viable cells in treatedrelative to nontreated conditions 1.

12、MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 The efficacy of the combination is also tested in vivo. A total of 3106 A549, H460, HCT116, or MDAMB231cells are injected s.c. into the flank region of 6-wk-old female athymic

13、 mice. When tumors reach 50 mm3, the2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEmice are randomized into four groups (n=7/group) for the following treatment: vehicle, RAF265 (12 mg/kgdaily), RAD001 (12 mg/kg daily), or both. All drug are administered over 14 d (6 d on, 2 d off, 6 d on), and the

14、drug combination is administered concurrently. Control mice receive the respective vehicles of both drugs.Animal weight and tumor volumes are taken twice weekly and expressed relative to initial tumor volume.Tumors are measured until achieving a relative volume of 10 times the initial volume, and th

15、e time to this endpoint is noted. Drug efficacy is assessed based on the tumor growth curve, growth delay, and tumor volumeinhibition percentage. The tumor growth curve is designed to depict the evolution of the relative tumor sizeover time. The tumor volume inhibition percentage (TVI%) is calculate

16、d 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Clin Sci (Lond). 2019 Apr 16;133(8):919-932. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Mordant P, et al. Dependence on phosphoinositide 3-kinase and RAS-RAF pathways drive the activity of RAF265, a novelRAF/VEGFR2 inhibitor, and RAD001 (Everolimus) in c